健康人外周血中A20基因3'UTR突变与多态性分析  

Characteristics of polymorphisms/mutations in 3'UTR of A20 gene in peripheral blood of healthy donor

作　　者：周玲玲[1,2] 朱丽花[3] 陆帅[1,2] 陈少华[2] 杨力建[2] 罗更新[4] 李扬秋[1,2,4] 

机构地区：[1]暨南大学再生医学教育部重点实验室,广东广州510632 [2]暨南大学血液病研究所,广东广州510632 [3]暨南大学附属第一医院风湿免疫科,广东广州510630 [4]暨南大学附属第一医院血液科,广东广州510630

出　　处：《暨南大学学报（自然科学与医学版）》2016年第1期7-11,共5页Journal of Jinan University(Natural Science & Medicine Edition)

基　　金：国家自然科学基金项目(91129720);广东省医学科研基金项目(A2014371;B2014223);暨南大学第一临床医学院科研培育专项基金青年项目(2015207)

摘　　要：目的:建立检测NF-κB负调控因子A20基因3'非编码区(3'UTR)单核苷酸多态性(SNP)和突变的方法和rs77191406的检测方法.方法:利用PCR扩增99名健康人外周血单个核细胞(PBMCs)DNA的A20基因3'UTR全长片段,并对PCR产物进行核苷酸序列分析.将所测A20 3'UTR序列与Gen Bank中的序列(NM_006290.3)通过BLAST程序对比,检出可能存在的基因突变.利用限制性片段长度多态性聚合酶链反应(PCR-RFLP)检测A20基因SNP rs77191406位点多态性.结果:在A20 3'UTR mRNA 3080位点检测出C>T突变,该突变在基因库中已有作为SNP登记(rs77191406),该SNP在7例(7.07%)健康人样本中检测到,但并未发现其他突变和多态性.成功建立PCR-RFLP检测A20基因SNP(rs77191406)的方法.结论:首次报道中国健康人群中A20基因3'UTR存在rs77191406,其意义有待进一步明确.Aim：To establish the method of identifying NF-κB negative regulatory factor-A20 gene 3′ untranslated region （3′UTR） polymorphisms and mutations, as well as and the method of identifying rs77191406. Methods： PCR and sequencing were used to identify A20 gene 3′ UTR region polymor- phisms in DNA samples of peripheral blood mononuclear cells （PBMCs） from 99 healthy individuals. Se- quencing was performed on the PCR products. The sequence of A20 gene 3′UTR was compared with the known sequence （NM_006290.3） in GenBank by the BLAST program and possible mutations were screened. Polymerase chain reaction-restriction fragment length polymorphism （PCR-RFLP） was used to identify rs77191406 polymorphism in A20 gene. Results： 3080 C 〉 T mutation was identified in A203′UTR mRNA locus, which has been registered as a SNP （rs77191406） in gene bank, and was identi- fied in 7 cases samples （7.07%） of healthy people. And we successfully establish PCR-RFLP to identify rs77191406. Conclusion：To our best knowlegde, it is the first identification of A20 gene 3′UTR SNP （rs77191406） in Chinese healthy individuals, the significance is needed to further investigation.

关 键 词：A20 3′非编码区 限制性片段长度多态性聚合酶链反应 单核苷酸多态性 

分 类 号：Q781[生物学—分子生物学] Q783.1